A plurality of oligonucleotides are caused to bind specifically to target nucleic acid which is predictive of a disease state or a biological condition within cells containing the target nucleic acid. The respective oligonucleotides, which may be functionilized or may be present in the form of oligonucleotide analogs, carry with them a plurality of synthons. Such synthons, which may be identiphores, toxiphores, or other precursors to biologically effective molecules, interact when specific binding of the respective oligonucleotides occurs at sites adjacent to each other on the target nucleic acid. The resulting interaction gives rise to the synthesis, generation or release of highly active biological molecules in situ in the cell in which the specific binding takes place. This permits the use of extraordinarily toxic molecules for use in killing cells containing the target nucleic acids. Imaging and other uses are also provided by the present invention.

The present invention is directed to the synthesis, generation or release of molecules within cells and tissues, which molecules have significant, biological effects. Such molecules are synthesized which permit the identification of the cells or tissues, which destroy the cells or tissues, which moderate, modulate, or enhance cellular or tissue function, or which otherwise have a diagnostic, therapeutic, nutritional or other biological effect.
The in situ generation of such molecules is specific to particular cells and tissues such that the molecules thus formed or released are delivered specifically to the cells or tissues in question. Such high specificity of delivery overcomes many therapeutic impediments and can provide high xe2x80x9cleveragexe2x80x9d with concomitant low side effects.
It has long been known to employ nucleic acid structures present in cells to serve as xe2x80x9ctargetsxe2x80x9d for diagnostic and therapeutic regimes. A classical example of such employment is evidenced by the field of antisense therapeutics. According to the antisense paradigm, a nucleic acid sequence or structure is identified as being associated with the production of a gene productxe2x80x94usually a peptidexe2x80x94which has deleterious effects upon cells or tissues. The target nucleic acid may be an oncogene, an mRNA associated with the genesis or development of a disease state or a hyperproliferative gene structures which proliferation is to be reduced or eliminated.
Under the antisense paradigm, oligonucleotides are designed which are specifically bindable to a nucleic acids sequence or structure, usually mRNA, the down regulation of which is desired. The oligonucleotide, usually in the form of a chemically modified analog or construct, is administered to the cells containing the targeted nucleic acid. The specific binding of the oligonucleotide interferes with expression of the mRNA, thus interfering with its function and expression. Protein normally expressed by the mRNA is either not expressed at all or is expressed in much lower quantity with a concomitant, beneficial therapeutic effect. The paradigm may also be used for diagnosis and research in ways which are well known to persons of ordinary skill in the art.
The antisense paradigm is well understood per se to persons of ordinary skill in the art and there are numerous review articles which describe in detail varying approaches to this practice.
Nucleic acids which can be used to target in accordance with some embodiments of this invention include mRNA molecules, which preferably have secondary structures such as stem-loop structures, or unique secondary structural sites, a such as the molecular interaction sites taught in U.S. application Ser. Nos. 09/076,440, 09/076,447, and 09/076,404, each of which is incorporated herein by reference in its entirety.
Potential target nucleic acids include, but are not limited to, mammalian, bacterial, fungal and viral mRNA. Oligonucleotides have been shown to bind to numerous targets including, for example, Candida (U.S. Pat. No. 5,691,461), protein kinase C (U.S. Pat. Nos. 5,703,054, 5,681,944, and 5,620,963), papillomavirus (U.S. Pat. Nos. 5,681,944 and 5,756,282), herpesvirus (U.S. Pat. No. 5,658,891), cytomegalovirus (U.S. Pat. Nos. 5,607,923 and 5,595,978), human immunodeficiency virus (U.S. Pat. No. 5,523,389), ras (U.S. Pat. Nos. 5,661,134, 5,582,986, and 5,576,208), Epstein-Barr virus (U.S. Pat. No. 5,242,906), cell adhesion molecules (U.S. Pat. Nos. 5,599,797 and 5,514,788), hepatitis virus (U.S. Pat. No. 5,576,302), Raf kinase (U.S. Pat. Nos. 5,654,284, 5,563,255, 5,656,612, and 5,744,361), p120 (U.S. Pat. No. 5,814,629), cell growth (U.S. Pat. No. 5,656,743), and multi-drug resistance associated protein (U.S. Pat. No. 5,510,239). Each of the above-identified patents is incorporated herein by reference in its entirety.
In addition to traditional antisense mechanisms, oligonucleotides have also been shown to act through mechanisms, for example, involving pseudo-half-knot formulations (U.S. Pat. No. 5,512,438), 5xe2x80x2-cap inhibition (U.S. Pat. No. 5,643,780), and triple helix formation (U.S. Pat. No. 5,834,185), each of which is incorporated herein by reference in its entirety. All such targets and many more may be used herein.
It has also been proposed to employ a pair of oligonucleotides to bind specifically with a mRNA in an antisense fashion. (see Viassov et al., xe2x80x9cBinary Systems of Oligonucleotide Conjugates for Sequence Specific Energyxe2x80x94Transfer Sensitized Photomodification of Nucleic Acidsxe2x80x9d NATO ASI Ser., Ser. C, 479 (DNA and RNA Cleavers and Chemotherapy of Cancer and Viral Diseases) 195-207, (1996). Viassov discloses that, along with one oligonucleotide there is a flourescent sensitizer, while the other oligonucleotide carries a structure which, when irradiated with ultraviolet radiation, is known to react with nucleic acids. Viassov demonstrates a rapid modification of the DNA to which the oligonucleotides were targeted when the two oligonucleotides are allowed to bind to adjacent sites on the DNA and be subsequently irradiated. The proximity of the photosensitizer to the group which transfers energy to the DNA was viewed to be important to the reaction with the DNA.
The technique of fluorescence resonance energy transfer (FRET) has been used, inter alia to identify point mutations in nucleic acids. A pair of oligonucleotides, each of which carries a fluorophores are caused to bind to nucleic acids. Through study of their fluorescence behavior, determination of the proximity of the fluorophores and, hence, the presence or absence of intervening base units, can give rise to the desired mutational information. FRET can also monitor ribozime interactions and seems to be useful generally in nucleic acid research.
Viassov et al in, xe2x80x9cSequence Specific Cleavage of Yeast tRNA-Phe with Oligonucelotides Conjugated to Diimidazole Constructionxe2x80x9d, Antisense Nucleic Drug Development, 7(1): 39-42, (1997), prepares oligonucleotides conjugated to a chemical construction having two histidine residues. Yurchenko et al, xe2x80x9cCleavage of Leishmania Minixe2x80x94Exon Sequence by Oligonucleotides Conjugated to a Diimidazole Constructionxe2x80x9d, Nucleosides and Nucleosides, 16 (7-9): 1721-1725, (1996), is directed to similar subject matter. While it has been known to conjugate a plurality of oligonucleotides to a single nucleic acid in a sequence specific fashion, all such techniques are believe to have been directed to the incapacitation or destruction of the nucleic acid target. While this can certainly have some useful applications, it is limited in its applicability.
It has been greatly desired to provide for the targeting of nucleic acids in a way different from the traditional, antisense approach. Thus, improvements over antisense methodology and over the methodologies for nucleic acid cleavage of, e.g. Viassov and Yurchenko are greatly desired. In particular, a methodology that does not necessarily involve the incapacitation of the target nucleic acid but which has a vastly more significant therapeutic potential have long been desired. The present invention is directed to this new paradigm of therapeutics and diagnostics.
The present invention provides a dramatic divergence in oligonucleotide therapeutics, diagnostics and related paradigms. In accordance with the invention, a nucleic acid target which has been identified as being present in the cell or tissue of interest is caused to be the object of specific binding by a plurality of oligonucleotides. By virtue of the specific binding of the two or more oligonucleotides, which binding is caused to take place at adjacent sites on the target nucleic acid, a molecular species is either synthesized, formed or released. The molecular species has great biological effect within the cell or tissue which, of course, carries the target nucleic acid. By virtue of the formation, synthesis or release of such biologically effective molecules, cells having the target nucleic acid may either be identified, imaged, killed, benefitted or modified in some significant way. Overall, this paradigm is denominated xe2x80x9cin situ binary synthesis,xe2x80x9d since a molecule having biological activity is created upon the specific binding of oligonucleotides to the target nucleic acid. The method is in situ since the formation, synthesis or release of the molecule takes place in situxe2x80x94within the cell or tissue.
It is apparent that the generation of these biologically active molecules, occurring as it does within the cell or tissue which has been targeted, is extraordinarily specific. Moreover, the generation of only a few molecules in this way can have an extraordinary and profound effect upon the cells or tissues. Thus, the molecules to be synthesized, generated or released are not spread out over an entire organism, but, rather, only within the cells or tissues which have been targeted. Extraordinary xe2x80x9cleveragexe2x80x9d can thus be obtained, such that a very small total amount of biologically active material need be formed in order to obtain a very large biological effect. Side effects are concomitantly diminished. Indeed, molecules may be selected in this way for generation, synthesis or release which could never be considered for diagnostic or therapeutic use under any other paradigm. As will be seen, even extraordinarily toxic molecules can find use in therapeutics so long as they are confined through the use of the present methodology to the particular cellular locus where they are needed.
The methodologies of the present invention do not necessarily involve modification of nucleic acid, or at least not the nucleic acid which serves as a targeting nucleic acid. Thus, unlike prior oligonucleotide diagnostic and therapeutic regimes, the objective of a specific binding of an oligonucleotide is not necessarily destruction or inhibition of the nucleic acid, usually mRNA, which has been targeted. Rather, the nucleic acid simply serves as the xe2x80x9clocator beaconxe2x80x9d and as a place for the docking of oligonucleotides by virtue of their specific binding. As will be seen, the ability of a plurality of oligonucleotides to specifically bind to a target nucleic acid at adjacent sites upon the nucleic acid permits the generation, synthesis or release of the biologically effective molecules in a very highly controlled fashion. Thus, wherever such specific binding at adjacent sites does not occur, generation, synthesis or release of the biological molecule does not take place. This high degree of control contributes to the utility of the present invention in that extraordinary toxic or potent molecules may be used without the worry of accidental generation or release of such molecules at locations other than the ones desired.
The present invention, to some extent, builds upon previous methodologies of oligonucleotide interaction with cellular nucleic acids, especially mRNA in the following way. It is well known to identify nucleic acids structures or sequences which are related to disease states. It is also known how to prepare oligonucleotides which have antisense sequences to the nucleic acids whose function or activity is to be moderated or ended. Moreover, a wide variety of chemical modifications of nucleic acids, and, indeed, a host of oligonucleotide analogs are known for this purpose. It is similarly well understood how to contact cells or tissues with an oligonucleotide or oligonucleotide analog in such a way as to cause formation of a heteroduplex between the oligonucleotide and the target nucleic acid. Such heteroduplex formation traditionally leads to the destruction of the nucleic acid or to a failure in its translation into protein. A number of intermediate results may attend this interaction. Thus, the binding of the oligonucleotide may activate RNAse H to cleave the nucleic acid. Alternatively, specific binding may simply make it impossible for the nucleic acid to be translated into protein. A number of other methodologies have been proposed for oligonucleotide interaction with nucleic acids, however these are generally attended by interference with the structure or activity of the target nucleic acid.
It follows from the foregoing that it is essential that the target nucleic acid be one which is important to disease states. It is not enough, under these prime paradigms, for the nucleic acid merely to be associated with a cell or tissue which is in a disease state. It may be immediately seen that interaction with a non-critical nucleic acid, such as an mRNA, even if very specific and entirely efficacious, will not lead to any beneficial results since the expression of the mRNA is not vital to survival of the cell or tissue. Moreover, antisense interaction with nucleic acids, while much more efficient than small molecule interaction with peptides, still requires the individual, specific binding of oligonucleotide to individual nucleic acids, such as mRNAs.
Contrarily, the present invention does not require the identification of a target nucleic acid which is vital to the function of a cell on which is causative of a disease state. Indeed the nucleic acid thus identified need not actually be translated into protein in large quantities within the cell or tissue in question. Rather, it is only necessary that the target nucleic acid be associated with or predictive of the existence of the disease state or other state of interest in cells or tissues to be treated. This is so because it is not the inactivation of the nucleic acid which is sought, but rather the use of that nucleic acid to target oligonucleotides to the cells or tissues which contain the target nucleic acid.
In accordance with preferred embodiments of this invention, a plurality of oligonucleotides are formulated to be specifically bindable to the target nucleic acid. Moreover, such oligonucleotides are formulated to have sequences which cause them to specifically bind to the target nucleic acid at adjacent sites. While it is possible that the oligonucleotides could bind at sites which are removed by one or two bases, it is greatly preferred that the oligonucleotides bind immediately adjacent to each other. It will, thus, be seen that the present methodology causes two oligonucleotides to enter a cell, tissue, or preparation having target nucleic acid and to bind to the target nucleic acid xe2x80x9cright next toxe2x80x9d each other. This close, predicable, and regulatable proximity of oligonucleotides to each other that contributes to the present invention.
Reliable and precise, specific binding of two designed oligonucleotides to each other within a cell containing a preselected, target nucleic acid permits the specific synthesis, formulation or release of a molecular species whenxe2x80x94but only whenxe2x80x94such specific binding occurs. Thus, a first one of the oligonucleotides carries with it a first portion of the biologically effective molecule to be synthesized, formulated or released. The second oligonucleotide carries with it a second portion of the biologically effective molecule. It is only when the first and second portions of the molecules are brought together in physical proximity and, preferably, with careful geometric alignment, that the biological molecule is either synthesized, generated, or released. It is also possible to have an essentially complete biologically effective molecule from part of one of the oligonucleotides with a moiety on the second oligonucleotide being capable of either altering, transforming, cleaving, or releasing the first biologically effective molecule or molecule precursor. In any event, whether the biologically effective molecule is actually synthesized through the joining together with covalent bonds of two or more fragments or synthons thereof, whether a precursor molecule is transformed, modified, or cleaved during this process, or whether a pre-formed, biologically effective molecule is cleaved or released from the oligonucleotides through the present methodology, the biologically effected molecule is not rendered effective unless and until the two oligonucleotides specifically bind at adjacent sites on the target nucleic acid within the cell, tissue, or in vitro preparation.
It will be seen that the first oligonucleotide carries with it a first synthon while the second oligonucleotide carries with or on it, or otherwise comprises, a second synthon. Persons of ordinary skill in the art will understand that the term xe2x80x9csynthonxe2x80x9d is extraordinarily broad and means a chemical moiety which is capable of interacting with another chemical moiety to give rise to chemical alteration.
While the invention is amenable to the use of a pair of oligonucleotides for interaction with the target nucleic acid, other paradigms are also possible within the spirit of the invention. Thus, an oligonucleotide may specifically bind to the target nucleic acid followed by the interaction of the heteroduplex with another species, which is not an oligonucleotide. Such other species may be, for example, a peptide, a carbohydrate or complex sugar, or a xe2x80x9csmallxe2x80x9d molecule, e.g. a non-oligomer. The confluence of two or more molecules in some specific relationship with target nucleic acid followed by the synthesis, generation or release of a molecular species having significant biological effect, especially toxic effect.
Certain aspects of the invention provide for the employment of two or more non-oligonucleotides in the practice hereof. Thus, specific binding of two or more non-oligonucleotide oligomers, carbohydrates or sugars, or small molecules may serve the purposes described herein. Thus, two or more small molecules could bind specifically and adjacently to the target nucleic acid, giving rise to a biologically active molecular species for, e.g. destruction of the cell containing the target nucleic acid or detection thereof.
The present invention provides for the detection of cells or nucleic acids found in the cells, with the killing of cells having target nucleic acids, and with certain other beneficial results. While, in all such cases, it may be seen that the respective oligonucleotides carry with them synthons, in accordance with the broad definition of that term, it may also be convenient to refer to the chemical moieties carried by the respective oligonucleotides in terms which are more nearly suited to their function. Thus, when it is desired to achieve the identification of cells or tissues containing target nucleic acid, the respective synthons carried by the oligonucleotides may be referred to as xe2x80x9cidentiphores.xe2x80x9d In this context, the identiphores are moieties which can interact with each other to give rise to either an interaction or a molecular species which can be detected in some way.
Detection may take place using any form of spectroscopy, x-ray examination, biochemical or biological analyses or otherwise. What is required is that the confluence of the two identiphores carried, respectively, by the oligonucleotides which specifically bind to the target nucleic acid, be detectable. The presence of the detectable moiety formed, synthesized or released by the conjunction in space of the two identiphores is, according to this embodiment, probative of the presence of the target nucleic acid in the cell, tissue or in vitro assay.
As persons of skill in the art will appreciate, x-ray detection may be employed. This is particularly beneficial when the identiphore contains a heavy metal such as a heavy metal contained within an heterocyclic structure such as a porphyrin, heme or similar species. The identiphores may form a charged transfer complex when in proximity with each other which complex may be detectable through spectroscopy. Such identiphores may also become covalently bonded with each other upon coming into proximity giving rise to a chemical moiety or molecule which can, itself be detected. Other forms of detection including magnetic resonance spectroscopy, proton magnetic resonance spectroscopy, electron resonance spectroscopy, fluorescence spectroscopy or electromagnetic spectroscopy may also be profitable employed in connection with this embodiment of the invention.
For example, the identiphores can comprise portions of a charge transfer complex of TTF-TCNQ, (Tetrathiafulvalene and Tetracyanoquinodimethane). Alternately, Bis-(ethylenedithio)dithiapyrene (ETDTYP) and Dibenzo barreleno tetracyano quino dimethane or Triphenylphosphine and Acrylonitrile could form such complexes. Other charge transfer complexes are known to persons of ordinary skill in the art. Thus, these molecular moieties may be caused to comprise the oligonucleotides which specifically bind to the target nucleic acid. Bringing the two identiphores into proximity gives rise to the detectable complex.
The use of identiphores for identifying the presence of target nucleic acid in cells or tissues may be used to diagnose certain cellular biological states such as disease states, especially hyperproliferative states and cancers.
As with most aspects of the present invention, the identiphores are best placed into spacial proximity with each other and also to have a preselected geometric relationship one to the other. This is especially true when intermolecular interactions are required such as charge transfer complexes and the like.
This aspect of the invention provides methods for identifying cells containing a preselected nucleic acid sequence. These methods comprise contacting the cells with a first oligonucleotide specifically bindable with the preselected nucleic acid, the first oligonucleotide comprising a first identiphore. The cells are also contacted with the second oligonucleotide specifically bindable with the preselected nucleic acid, preferably at a site immediately adjacent to the site where the first oligonucleotide has bound to the nucleic acid. The second oligonucleotide also comprises an identiphore. The first and second identiphores are detectable when in proximity with each other. Thus, they may either react with each other to form a new molecular species, cause a species to be released from the oligonucleotides for detection, or may give rise to an entirely new moiety which, itself, may be detected in any of the ways known to persons of ordinary skill in the art.
In accordance with other preferred embodiments of the present invention, the synthons carried by the respective oligonucleotides may give rise to toxic molecules. Such molecules may either be synthesized, generated or released upon the specific binding of the two oligonucleotides to adjacent sites on the target nucleic acid. The generation of a toxic molecule as a result of this specific binding gives rise to cell death and possibly even destruction of tissue material in adjacent cells. It will be appreciated that many highly toxic molecules exist in nature and otherwise which can effectuate widespread destruction on a cellular and tissue level. Prior therapeutic regimes have never been able to exploit this phenomenon, since systemic or untargeted toxicity has never been able to be avoided heretofore. The present invention, however, permits this.
Thus, the present invention provides methods of killing cells, which cells contain a preselected nucleic acid. The method comprises contacting the cells with a first oligonucleotide specifically bindable with the selected preselected nucleic acid. The first oligonucleotide comprises a first toxiphore. It would be understood that the term xe2x80x9ctoxiphorexe2x80x9d is meant herein to mean a chemical species which, when combined with one or more additional toxiphores, gives rise to a toxic molecular species, interaction or phenomenon. Such toxicity is to be expressed on a cellular or tissue level; it is not necessary that any particular nucleic acid be inactivated or xe2x80x9ckilledxe2x80x9d thereby. The method further comprises contacting these preselected nucleic acid sequence with a second oligonucleotide specifically bindable with it. The second oligonucleotide contains a second toxiphore. The first and second toxiphores either synthesize, generate or release a toxin upon coming into proximity with each other as is the case when the two oligonucleotides, carrying their toxiphores, specifically bind to adjacent sites on the target nucleic acid.
It is, of course, preferred that the toxiphores be placed into spatial proximity when the oligonucleotides specifically bind to the target nucleic acids. Additionally, it is greatly preferred that the toxiphores be oriented geometrically so as to facilitate the formation of the molecular species, its generation or release.
In accordance with preferred embodiments, the toxiphores become chemically bonded one to the other to give rise to the toxic, molecular specie. Such specie is, of course, highly biologically effective. It is also within the spirit of this invention that one toxiphore modify the second toxiphore to give rise to the biologically effective, toxic molecule. One toxiphore may also cleave from the oligonucleotide, the heteroduplex, or otherwise, a toxic species.
As will be appreciated, it is desired that the preselected nucleic acid, to which the oligonucleotides and their related toxiphores will specifically attach, be predictive of a cellular biological state. Such state is usually a disease state, especially a hyperproliferative state or one indicative of cancer. Other cellular or tissues states may also be associated with the preselected nucleic acid and, indeed, the present invention may be used in the sense of a probe as well as in the sense of a therapeutic or diagnostic. A host of research functions may also be benefitted through use of the present invention.
The toxic molecule which, in accordance with preferred embodiments of this invention, are synthesized, generated or released may be anything which has effective, preferably powerful cellular toxicity within related tissues or organs. For example, the toxin may be a peptide such as ricin, sarcin, or diphtheria, botulism, etc. toxins. In such cases, the toxin would comprise a functional domain of the toxin. The toxin may be bacterial. The toxin need not be a peptide. For example, the toxin could be an amphibian toxin such as tetradotoxin or bufotoxin. The toxins may also be from invertebrates, such as arachnids, or from reptiles or amphibians. In short, any number of toxins may be employed in conjunction with the present invention. It is preferred that the toxin be very powerful such that only a few, perhaps as few as one or two molecules of the toxin are necessary to effect cell death. Numbers of toxin molecules fewer than ten are preferred in accordance with certain embodiments to kill any given cell. As will be appreciated, a preferred embodiment of the present aspect of the invention is for the treatment of disease state in mammals. Thus, a tissue of a mammal is contacted with the oligonucleotides having toxiphores associated therewith under conditions such that heteroduplexes can be formed. Alternately, in vitro embodiments, heteroduplex formation is encouraged through appropriate selection of reaction conditions. Upon the specific binding of the oligonucleotides to the preselected nucleic acid, the toxiphores interact with each other to give rise to a toxic molecule or other toxic effect.
When the toxin is a peptide, it can be sufficient that a carboxylic acid xe2x80x9cendxe2x80x9d of a toxiphore be brought into spatial proximity with an amine xe2x80x9cendxe2x80x9d of another toxiphore, and the same be carefully oriented in space and with geometric precision. In such case, the activation energy required for the formation of the peptide bond may be significantly lowered such that spontaneous generation of the peptide bond occurs. The resulting, toxin is then either active immediately or can be released or cleaved from the complex for toxic activity. Organic molecules other than peptides may also be prepared in this way, cleaved, modified, or released.
The chemical species which effects the toxic activity may also be other than an organic species; they may be organometallic or even inorganic. Thus, one toxiphore might contain a complex metal ion which, in uncomplexed form or in a different oxidation state is highly toxic. The interaction with the second toxiphore may give rise to the toxic form of the metal ion or to its complex. A number of xe2x80x9cheavyxe2x80x9d metallic species and some which are not so xe2x80x9cheavyxe2x80x9d are known to be highly toxic in one or another form. All of these are contemplated by the present invention.
As is apparent, the present invention provides for the in situ synthesis of chemical species. This is accomplished through specifically binding first and second oligonucleotides to a preselected nucleic acid at adjacent sites of the nucleic acid. First and second sythons, related to or forming part of the respective oligonucleotides, are then reacted with each other or caused to interact to give rise to a new molecular species or an altered form of an existing molecular species. Significant biological effects may thus be generated upon the cell or tissues comprising those cells.
As will be apparent, a wide variety of organic, organometallic and other inorganic molecules can be formed through the practice of the present invention. As discussed herein above, peptides may be prepared by bringing together two synthons comprising peptide residues, which residues, when carefully oriented in space and oriented as to geometry may form the peptide bond either spontaneously or through the intervention of a catalyst. Indeed, a catalytic agent may also be included in one or more of the oligonucleotides. A host of other organic molecules may similarly be formed as will be apparent to persons of ordinary skill in the art. Organometallic materials may also be prepared hereby and the oxidation state of organic, and organometallic species may be changed through intermediation of oxidizing and reducing agents forming part or all of one or both of the synthons. The charged transfer complexes as discussed herein before may also be placed into apposition and may be used in either a catalytic, synthetic, biologically active or other functional applications. As is apparent, the ability of the present invention to orient synthons in precise proximity and in exact geometric relationship one with the other permits the overcoming of activation energies which would otherwise apply. Accordingly, even reactions which seem extraordinarily slow under solution conditions, may run at sensible rates when synthesized in accordance with the present invention.
It is also within the spirit of the present invention to alter, cleave, release or otherwise furnish chemical moieties through the interaction with the plurality of synthons placed into proximity in this way. Thus, active chemical species may be cleaved from the parent oligonucleotides and from the heteroduplexes to which they have been attracted through the action of the other synthon or otherwise.
Accordingly, the present invention provides for the preparation of such chemical species in amounts which are detectable in cells containing the target nucleic acid. It is, of course, preferred that such nucleic acid be predictive of a biological condition or disease state such that the preparation and delivery of the molecules thus formed or furnished is delivered in situ to such cells. Diagnosis, therapy, or killing of the cell may ensue.
The present invention also provides compositions of matter. In accordance with one embodiment, the invention provides a pair of oligonucleotides each of which is specifically bindable with a preselected nucleic acid. The binding of the pair of oligonucleotides is preferably at adjacent sites on the preselected nucleic acid. Each of the oligonucleotides comprises a portion of a molecular species, which species is formed when the adjacent, specific binding of the oligonucleotides to the nucleic acid occurs. Any of the foregoing methodologies may give rise to pairs of oligonucleotides which are contemplated by the present invention. Indeed, the present invention may also be applied to additional oligonucleotides with additional synthons (toxiphores, identiphores, etc.) whereupon binding of three or more oligonucleotides to adjacent sites on the target nucleic acid is accomplished with results analogous to those described herein above. As will be appreciated, the oligonucleotides may give rise to toxins, charge transfer complexes, detectable molecules for detection and sensing, cell regulatory moieties, nutritional species, and to a wide variety of other compounds having biological activity. The present invention is also directed to compositions of matter comprising a pair of oligonucleotides as herein above together with a diluent or carrier, especially a pharmaceutically acceptable diluent or carrier.
In the present description of the invention, the term oligonucleotide has been used very broadly. Persons of ordinary skill in art will appreciate that the term includes wild type oligonucleotides, those which are unmodified in any way in terms of their chemical construction or substituents. The term also includes semisynthetic and modified oligonucleotides and molecules which are analogous to oligonucleotides. In this context, oligonucleotide includes peptide nucleic acids, xe2x80x9cPNAsxe2x80x9d, molecules which are analogous in function and spatial relationships to nucleic acids and which may be used in oligonucleotide therapeutics such as antisense and the like. These have been well-characterized in the literature and are set forth in number of U.S. patents, many of which are assigned to the assignee of the present application. Each of these are incorporated herein by reference.
Thus, numerous other identophores have been shown to bind mRNA including, for example, peptide nucleic acids (U.S. Pat. Nos. 5,641,625, 5,700,922, 5,719,262, 5,714,331, 5,766,855, 5,773,571, 5,786,461, and 5,736,336, each of which is incorporated herein by reference in its entirety), ribozymes (U.S. Pat. Nos. 5,599,706, 5,801,158, 5,639,655, 5,635,385, 5,599,704, 5,610,052, 5,766,942, and 5,747,335, each of which is incorporated herein by reference in its entirety), and small molecules (U.S. Pat. Nos. 5,668,165, 5,641,486, and 5,683,873, each of which is incorporated herein by reference in its entirety).
Chemically modified oligonucleotides include those which are modified in the backbone, e.g. phosphorothioate, methylphosphonates and a host of other backbone modifications as well as modifications to the substituents on the sugar rings present in the oligomers. Further modifications may be had on the base structure in a number of ways. These are exemplified by a number of U.S. patents including many owned by the assignee of the present application. Each of these in incorporated in full by reference in order to provide more explanation of the full scope of the oligonucleotides which may be employed in connection with the present invention: U.S. Pat. Nos. 5,618,704, 5,608,046, 5,792,844, 5,808,023, 5,834,607, 5,138,045, 5,378,825, 5,359,051, 5,212,295, 5,736,294, 5,614,617, 5,670,633, 5,681,941, 5,359,044, 5,210,264, 5,218,105, 5,610,289, 5,506,351, 5,391,667, 5,514,786, 5,489,677, 5,541,307, 5,386,023, 5,457,191, 5,578,718, 5,644,048, 5,623,065, 5,719,271, 5,637,684, 5,783,682, 5,539,082, 5,614,621, 5,459,255, 5,521,023, 5,539,083, 5,510,476, 5,629,152, 5,554,746, 5,571,902, 5,519,134, 5,543,507, 5,734,041, 5,506,212, 5,714,606, 5,717,083, 5,831,014, 5,677,437, 5,623,070, 5,688,941, 5,808,027, 5,750,692, 5,639,873, 5,731,438, 5,798,360, 5,646,265, 5,760,202, 5,587,361, 5,635,488, 5,587,469, 5,587,420, 5,770,713, 5,214,597, 5,817,489, 5,811,534 and 5,602,240.
While the present invention has been exemplified in terms of the use of a plurality of oligonucleotides to bind specifically to target nucleic acid and to effect the synthesis, generation or release of biologically useful molecules thereby, certain other materials may also be employed. Thus, it may be seen that a first oligonucleotide carrying a synthon (identiphore, toxiphore, etc.) may be caused to bind specifically to a site of a target nucleic acid. Another biooligomer may then be caused to bind to the heteroduplex thus formed. This latter oligomer need not necessarily be an oligonucleotide. Thus, certain peptides are known to be able to complex with heteroduplexes formed between a nucleic acids and oligonucleotides. It is possible to include a synthon along with such peptide and to cause the same to bind the heteroduplex thus bringing the synthon of the peptide and the synthon carried by the oligonucleotide into proximity with the results explained above. Many other variations are possible in accordance with the full spirit of the present invention.
It is also believed to be possible to use a single oligonucleotide to accomplish the function presently exemplified through a plurality of oligonucleotides. Thus, a single oligonucleotide having two portions, each of the two portions carrying a synthon (identiphore, toxiphore, etc.) may be employed. If the two portions of the single oligonucleotide can bind at adjacent locations on the target nucleic acid, thus bringing the synthons into proximity, the spirit of the present invention may be accomplished. It may also be that a single oligomer which first binds with the target nucleic acid and then binds with the heterodymer formed by the first part of the oligomer and the target nucleic acid could be used in conjunction with certain embodiments of the present invention.

In situ binary synthesis of biologically effective molecules


A method for playing a gaming device where selected bets enable various bonuses. In the preferred embodiment 5 different bonuses are offered depending on credit per line bets, and said bonuses may occur in combination.

Gambling devices, or gaming devices including slot machines, allow players to wager something of value in the hopes of winning something of greater value. Originally slot machines were mechanical devices employing 3 physical reels with various symbols painted or affixed to them. Upon inserting a coin and pulling a spring-loaded handle, the reels were set in motion and players were paid or not depending upon where the reels stopped and which symbols were displayed across the display portion of the machine.
Slot machines have evolved greatly since the original gaming devices discussed above. Most slot machines in use today are electronic computers, and the symbols are displayed in video form. Players often prefer video display slot machines over the traditional slot machines using mechanical reels. The video platform also offers more flexibility in development for manufacturers. The common term today for a gambling machine is a gaming device, which is used herein to include gambling devices such as slot machines, video poker, and other gambling games whether reel, video or otherwise.
Newer video slots typically display 5 reels side-by-side that spin on a common axis instead of the traditional 3 (or more) mechanical reels. These newer video slots usually show 3 stopping positions of each reel yielding a visible matrix of 3 rows by 5 columns. Sometimes the number of reels and reel positions displayed vary.
Players typically wager on 1 or more paylines that run in different paths through the reel positions displayed. Unlike newer models of gaming devices, early slots paid only for matching symbols straight across the center (a single payline). Traditionally the path of each payline takes 1 adjacent position of each reel, so on a 5-reel game the payline is usually 5 positions in length. However, today these payline paths may number in the hundreds, they may take any path, and they may not always span adjacent reels nor even be on contiguous reel positions. Players may wager on multiple paylines and may even wager multiple credits per payline. Reel symbols occurring in various combinations on the paylines are compared to a schedule of winning events commonly called a paytable to determine a win or loss. Often there are wild symbols that substitute for other symbols, and symbol combinations that trigger a bonus or feature game. Scatter pays are also common where certain symbol(s) pay anywhere in the visible display, and they don't have to be on any payline. Wins are usually rewarded with monetary awards from a coin hopper provided in the machine or tickets redeemable for cash.
Second screen features (or more) are common. A "second-screen" bonus game is usually separate and distinct from the normal reel display, and a player might select a car in a car race or scratch from a selection of video lottery tickets to earn credits, free games or anything of value. Some games even offer third screens or more, enhancing player interest and intrigue. These bonus game features typically occur randomly every 50 plays or so and are embedded in the game mathematics. Their occurrence is not conditioned upon an additional bet.
Some games let creative players play consecutive games without manual intervention. Sometimes players wedge a toothpick or folded matchbook cover in such a manner as to keep the play button depressed. Provided they have sufficient credits, consecutive games play off by themselves. Although these players have basically fashioned an autoplay device, each game is still a single, discrete event that requires a separate wager. However, in newer gaming devices a "game" might be comprised of multiple slot machines spins of a traditional nature plus other interactions and events.
In general, the more interactive a gaming device is, the more entertaining it is, and the greater its entertainment value the more players will play it. Accordingly, more play normally translates to greater profits for gaming device operators.
Other types of gaming devices offer side bets. For example, as disclosed in U.S. Pat. No. 6,283,474 to de Keller, players play blackjack, but may wager separately who gets closest to a "21" point score. Similarly, in U.S. Pat. No. 6,019,374 to Breeding, a gaming device is provided which offers a side bet to participate in a super jackpot game.
Side bets have also been around for a very long time. Aristocrat, a major slot machine manufacturer headquartered in Australia, makes games that offer side bets embedded within a video slot machine. Their Cashman series offers bonus features that can only be achieved when a side bet is made. This side bet is only offered when the player has first bet the maximum number of lines offered on the machine. For example, after a player bets the maximum 20 paylines they are offered the chance to make a side bet that costs the equivalent of betting another 5 paylines. Thus, these games entice the player to raise their bet. Since casinos and operators normally retain a percentage of the bet, this side bet usually results in greater revenues and profits. While this is a somewhat creative means to offer a side bet, it is still just an additional bet. Side bets are separate wagers on separate events.
Some gaming devices offer multiple bets. Triple play poker as described in U.S. Pat. No. 5,823,873 to Moody allows a player's held cards to be played as multiple poker hands with each draw performed independently, usually resulting in 3 different outcomes. Each additional hand requires an additional wager that can become expensive, since it costs 3 times as much to play.
Regardless of the type or form of gaming device—whether reel slot, video game, some other type or combination—the basic method of wagering has remained largely the same for years. The player inserts coins or otherwise obtains credits on a machine, commits a wager, plays the game and then is paid or not depending on the outcome.
Slot machines today play in many different denominations. Credits on one machine might be worth 1 cent (or less), while others are worth $100. Some machines today even allow you to select the denomination within the machine and to change it between games. One advance in slot machine development was the use of a credit meter. With a credit meter, a player could insert more coins than were needed to play a single game, and thus have a pool of funds to draw upon. Ten nickels inserted would yield ten credits, for example. Then, the player could play one game that required ten nickel credits, ten games that required one nickel credit each, or anything in between. The use of a credit meter also allowed winnings to be accumulated on the machine, instead of always being paid out in coins each time the player won. A player could rack up credits and then choose to cash out at their leisure by the use of a special button on the machine for this purpose.
In the 1980s and 1990s, slot machine manufacturers began adding currency acceptors to their machines. Players could obtain machine credits by simply inserting paper currency. In recent years, manufacturers have even added devices that could dispense currency instead of, or in addition to, coins. (These are known as note hoppers and operate similar to bank cash machines or ATMs). Many new machines pay players in paper tickets or scripts that may be redeemed elsewhere or reinserted into similar machines that read and accept such paper. These are often called ticket-printer machines. Some machines may even accept credit cards or other cards that have value.
A traditional video slot machine commonly has 2 rows of bet buttons plus a service and cashout button. The top row dictates the number of paylines, often from 1 to 9 or more. (In the case of the Cashman example cited earlier, the top row also contains the side bet button.) The bottom row has the credit per line bet buttons, which are commonly from 1 to 10. These limits change depending on the game. A typical one-cent game today may offer up to 20 paylines and 20 credits per line for a total bet of 400 credits or $4.00 per play, or more. Play is commonly initiated by pressing the credit per line button, which is multiplied by the number of currently selected paylines to arrive at the total bet.
As a promotion, casinos sometime configure certain slot machines for tournament play. Slot tournaments are player versus player competitions administered by casino staff. Players gain entry through a variety of means, such as achieving VIP status in the casino's players club, paying an entry fee or simply for signing up. In these tournaments, players do not wager anything directly or at all, but person(s) with the highest credit scores receive something of value from the casino. Since there is no wager, the slot machines are set to a free play mode where the goal is to get more credits than the other players. Players are not paid directly for credits earned. Tournaments are marketing programs that rely on gaming devices for implementation.
Time on a particular device is a huge factor in the gaming industry today. Time on device is important not only for direct profits to gaming device operators, but also indirectly. The more time spent in one gambling property means more profit opportunities for the casino. Restaurants, shows, gift shops, hotel rooms, etc., all give the casino the chance for more profits. In general, the more time a player spends in a gaming establishment, the greater the likelihood they will return and spend even more. Operators strive to keep you in their establishment, which generally means more profits, and typically provide numerous incentives such as free or inexpensive food and drinks specifically for this purpose.
Marketing studies have revealed that players do not mind losing so much, as long as they can have a good time playing. Most casino gamblers don't really expect to win, but they do expect to play for a reasonable amount of time. This is consistent with newer trends in gaming where gambling for the typical person is more of an entertainment experience. With the proliferation of casinos in America in recent years, casino gambling has become mainstream entertainment. Perhaps the most important part of that entertainment value is "time on device", or how long you get to play for your money. Time on device is critical to a positive gambling experience.
Accordingly, recent attempts have been made to ensure greater time on device for players. Perhaps the most common method today on slot machines is to offer more winners of lesser amounts. To this end slot makers design games with a greater mathematical win frequency and a reduced paytable. Especially in newer video slots these win or hit frequencies reach 50% or more. In practice, this means you might bet 10 coins per line on 9 paylines (90 coins total) only to win 20 coins. Even though this is clearly a net loss for the player, the 20 is still touted as a win. This trickling back of credits to the player takes their money more slowly, recognizes them as winners (even if they're losing) and extends their playtime for a given amount of money to bet.
One problem with high hit frequencies is that player returns become meaningless. While time on device is generally lengthened, batting 90 to win 20 eventually becomes tedious and boring. Players soon realize that in these types of gaming products, they are not really winning, even if the machine displays "winner", but rather—they have simply lost less. The reduced payable means the allure of big winners is diminished. The tradeoff of more common winners is smaller winners.
Other enticements also keep players playing longer. As mentioned occasional bonus features or second-screen events that award free plays or credits are common ways to motivate players. Random payouts and mystery jackpots (typically paid anywhere on a bank of networked gaming machines) are also popular. Progressive awards, where certain outcomes pay an amount that increases with credits played until won, also give players something special to play for. (Qualifying for progressive awards is often conditioned on players first wagering the maximum allowable bet.)
Some less common gaming devices employ a hold and re-spin feature. In these games the reels are spun once, then desirable symbols are held similar to draw poker and the rest are spun again in order to make more favorable combinations. Still other esoteric and non-traditional concepts let players buy a predetermined amount of time and play without even being present as in U.S. Pat. No. 6,077,163 to Walker.
Different jurisdictions impose different regulations onto casino operators and gaming machine manufacturers. For example, riverboat casino legislation in Mississippi resulted in casinos being built in the mud that don't even float. These riverboats don't navigate and can't even move. In other markets, however, riverboat casinos are required to cruise some distance periodically to qualify under the regulations.
Similarly, different states place different requirements onto gaming devices. One common rule requires a skill component. Thus in some markets video draw poker games are the standard. Skill is needed to make the best choice as to cards retained and cards replaced on the "draw."
Some states don't allow traditional slot machines, but only bingo or lottery games that resemble slot machines. These games, often called "Class II" games, usually require another button press or two to set the reels spinning. Commencing with the bet, a bingo card is selected. The next button push selects a series of bingo balls, and a win or loss outcome is determined in accordance with the rules of bingo. The next press sets slot machine reels spinning and the resulting outcome is that commensurate with the bingo game. As an example, a bingo winner paying 10 is represented as a slot machine result paying 10. Thus clever manufacturers have fashioned a bingo game simulating a slot machine that is legal in some markets.
Still other markets require "central determination." Unlike traditional slot machines that stand alone independent of each other, centrally determined gaming devices are networked to a central server. This server dishes random numbers or gaming device outcomes to each device on the network. Thus a central accounting point may be maintained in a government office that controls each device statewide. While imposed by regulation, this also helps with accounting controls, general security and the verification process over large jackpot payouts. These are sometimes called video lottery games. Many other variations exist on gaming devices and the systems that work with them.
As indicated lower denomination games have become very popular recently. In the early 1990s games were generally nickel, quarter and dollar denomination. Today one-cent games are among the most popular. Since the denomination has fallen, so have average bets. Thus, manufacturers and gaming device operators are striving for ways to keep the bets high to maintain profitability. To these ends, some gaming device makers incorporate elaborate second- and third-screen bonuses with multiple bonus features embedded within their video slots. But, irrespective of the wagering method, games that are too complicated are generally less successful, since most players prefer new games that are easy to learn and play. So, it is a balancing act to keep games interesting and engaging, but simple. Likewise, keeping the bets high enough to return traditional or better profits while offering lower denominations is also a major objective of game designers today.


Care agents based on matrix particles having a perfume component and washing-active surfactant components can be used in body-cleansing compositions and household cleaners.

The invention relates to care agents, which comprise matrix particles having a perfume component and a washing-active surfactant component, wherein the matrix particles preferably consist of a gelatinous polysaccharide. The care agents can be used, for example, in body-cleansing compositions and household cleaners.
The encapsulation of products in various technical fields (e.g. medicaments, dyes, foods etc.) is known in order to protect products from undesired influences prior to use, and to use them in a targeted manner.
In the field of care agents, it is known to encapsulate surfactant components and optionally also perfume components as coacervates. Coacervates contain a shell (e.g. made of gelatin, cellulose, gum arabic, polyvinyl alcohol and others, or combinations thereof) and a filling material.
Release of the filling material from the coacervate is only possible by targeted mechanical or physical destruction of the shell. Easy release of the filling material associated with the use is desirable.
As a result of the precipitation processes, loading of the coacervate capsules with relatively large solid or water-soluble constituents is not possible. However, for many application forms, e.g. the uptake of colored particles, this is desired.
An object of the present invention was to protect care agents, in particular, for body-cleansing and technical use, from undesired impairment prior to use, to be largely unlimited in the choice of ingredients, and to permit easy release of the ingredients upon use.
We have found care agents, which comprise matrix particles having a perfume component and a washing-active surfactant component.
The care agent according to the present invention requires the action of only a small force to release the perfume component and optionally, further ingredients. A further unexpected advantage of the matrix particles according to the present invention is that, in contrast to coacervates, they can also be loaded with solid or water-soluble constituents. In the case of coacervates, this is not possible because of the filling process.
Matrix particles for the care agents according to the present invention are known per se (WO 98/15192, Gellan Gum, A Multi-functional Polysaccharide for Gelling and Texturising, Monsanto company brochure from 1992, Technical Bulletin xe2x80x9cThe Preparation of Kelcogel(copyright) Gellan Gum Productsxe2x80x9d company brochure from The Nutra Sweet Kelco Company RC-137).
The matrix particles for the care agents according to the present invention are multifunctional polysaccharides based on starch, guar, carob seed grain, tragacanth units, xanthan gum, gum arabic, carboxymethylcellulose, alginates, methylcellulose and karaya gum. Particular preference is given to polysaccharides made from glucose, glucuronic acid and rhamnose. Preference is given here to a building block ratio of 2:1:1.
More preferred matrix particles for the care agents according to the present invention contain gellan gum. Gellan gum is a water-soluble polysaccharide obtained by aerobic fermentation from Pseudomonas eludea. The microorganisms are supplied by a nutrient medium with a carbon source, phosphates, organic and inorganic nitrogen compounds and trace elements. Prerequisites for growth conditions are sterile working procedures, the introduction of oxygen, agitation, and temperature and pH control. The fermentation mixture is then pasteurized in order to kill the living cells. Gellan gum is obtained from the fermentation mixture.
During the addition reaction of salts onto the carboxyl groups, aggregation of the matrix particles takes place, X-ray diffraction analysis showing that the matrix particles have the structure of a laevorotating parallel double helix, which is folded three times.
Matrix particles having a perfume component can be prepared, for example, by dissolving a multifunctional polysaccharide in water at a suitable hydration temperature, adding the perfume component and preparing an emulsion, the emulsion being added dropwise to an aqueous solution of salts.
Polysaccharide and perfume component are generally used in the ratio 1 to 30, preferably 1 to 20.
Salts are preferably calcium chloride, potassium chloride, magnesium chloride and sodium chloride.
The aqueous solution generally comprises 0.001 to 2 parts by weight, preferably 0.01 to 0.5 parts by weight, of salts.
The size of the matrix particles can be controlled by the size of the dropping capillary.
The care agents according to the present invention generally comprise 0.1 to 20 parts by weight, preferably 5 to 15 parts by weight, more preferably 8 to 12 parts by weight, of matrix particles.
The perfume component can contain one or more, preferably 20 to 60, fragrances.
The perfume component is essentially insoluble in water.
Examples of fragrances for the care agents according to the present invention are given, for example, in S. Arctander, Perfume and Flavor Materials, Vol. I and II, Montclair, N. J., 1969, published privately, or K. Bauer, D. Garbe and H. Surburg, Common Fragrance and Flavor Materials, 3rd. Ed., Wiley-VCH, Weinheim 1997.
Individual examples which may be mentioned are:
Extracts from natural raw materials such as essential oils, concretes, absolutes, resins, resinoids, balsams, tinctures, such as, for example, ambergris tincture; amyris oil; angelica seed oil; angelica root oil; aniseed oil; valerian oil; basil oil; wood moss absolute; bay oil; mugwort oil; benzoin resin; bergamot oil; beeswax absolute; birch tar oil; bitter almond oil; savory oil; bucco leaf oil; cabreuva oil; cade oil; calmus oil; camphor oil; cananga oil; cardamom oil; cascarilla oil; cassia oil; cassia absolute; castoreum absolute; cedar leaf oil; cedarwood oil; cistus oil; citronella oil; lemon oil; copaiva balsam; copaiva balsam oil; coriander oil; costus root oil; cumin oil; cypress oil; Davana oil; dill herb oil; dill seed oil; eau de brouts absolute; oakmoss absolute; elemi oil; estragon oil; eucalyptus citriodora oil; eucalyptus oil; fennel oil; spruce needle oil; galbanum oil; galbanum resin; geranium oil; grapefruit oil; guaiac wood oil; gurjun balsam; gurjun balsam oil; helichrysum absolute; helichrysum oil; ginger oil; iris root absolute; iris root oil; jasmine absolute; calamus oil; camomile blue oil; camomile Roman oil; carrot seed oil; cascarilla oil; pine needle oil; spearmint oil; caraway oil; labdanum oil; labdanum absolute; labdanum resin; lavandin absolute; lavandin oil; lavender absolute; lavender oil; lemongrass oil; lovage oil; distilled lime oil; pressed lime oil; linaloe oil; litsea cubeba oil; bayleaf oil; mace oil; marjoram oil; mandarin oil; massoi bark oil; mimosa absolute; musk seed oil; musk tincture; clary sage oil; nutmeg oil; myrrh absolute; myrrh oil; myrtle oil; clove leaf oil; clove flower oil; neroli oil; olibanum absolute; olibanum oil; opopanax oil; orange flower absolute; orange oil; origanum oil; palmarosa oil; patchouli oil; perilla oil; peruvian balsam oil; parsley leaf oil; parsley seed oil; petitgrain oil; peppermint oil; pepper oil; pimento oil; pine oil; pennyroyal oil; rose absolute; rosewood oil; rose oil; rosemary oil; dalmation sage oil; Spanish sage oil; sandalwood oil; celery seed oil; spike lavender oil; Japanese anise oil; styrax oil; tagetes oil; fir needle oil; tea-tree oil; turpentine oil; thyme oil; Tolu balsam; tonka absolute; tuberose absolute; vanilla extract; violet leaf absolute; verbena oil; vetiver oil; juniper oil; wine lees oil; absinthe oil; wintergreen oil; ylang oil; hyssop oil; civet absolute; cinnamon leaf oil; cinnamon bark oil; and fractions thereof, or ingredients isolated therefrom;
individual fragrances from the group of hydrocarbons, such as, for example, 3-carene; xcex1-pinene; xcex2-pinene; xcex1-terpinene; xcex3-terpinene; p-cymene; bisabolene; camphene; caryophyllene; cedrene; farnesene; limonene; longifolene; myrcene; ocimene; valencene; (E,Z)-1,3,5-undecatriene;
of aliphatic alcohols, such as, for example, hexanol; octanol; 3-octanol; 2,6-dimethylheptanol; 2-methylheptanol, 2-methyloctanol; (E)-2-hexenol; (E)- and (Z)-3-hexenol; 1-octen-3-ol; mixture of 3,4,5,6,6-pentamethyl-3/4-hepten-2-ol and 3,5,6,6-tetramethyl-4-methyleneheptan-2-ol; (E,Z)-2,6-nonadienol; 3,7-dimethyl-7-methoxyoctan-2-ol; 9-decenol; 10-undecenol; 4-methyl-3-decen-5-ol; of aliphatic aldehydes and 1,4-dioxacycloalken-2-ones thereof, such as, for example, hexanal; heptanal; octanal; nonanal; decanal; undecanal; dodecanal; tridecanal; 2-methyloctanal; 2-methylnonanal; (E)-2-hexenal; (Z)-4-heptenal; 2,6-dimethyl-5-heptenal; 10-undecenal; (E)-4-decenal; 2-dodecenal; 2,6,10-trimethyl-5,9-undecadienal; heptanal diethyl acetal; 1,1-dimethoxy-2,2,5-trimethyl-4-hexene; citronellyl oxyacetaldehyde;
of aliphatic ketones and oximes thereof, such as, for example, 2-heptanone; 2-octanone; 3-octanone; 2-nonanone; 5-methyl-3-heptanone; 5-methyl-3-heptanone oxime; 2,4,4,7-tetramethyl-6-octen-3-one; of aliphatic sulfur-containing compounds, such as, for example, 3-methylthiohexanol; 3-methylthiohexyl acetate; 3-mercaptohexanol; 3-mercaptohexyl acetate; 3-mercaptohexyl butyrate; 3-acetylthiohexyl acetate; 1-menthene-8-thiol;
of aliphatic nitriles, such as, for example, 2-nonenenitrile; 2-tridecenenitrile; 2,12-tridecenenitrile; 3,7-dimethyl-2,6-octadienenitrile; 3,7-dimethyl-6-octenenitrile;
of aliphatic carboxylic acids and esters thereof, such as, for example, (E)- and (Z)-3-hexenyl formate; ethyl acetoacetate; isoamyl acetate; hexyl acetate; 3,5,5-trimethylhexyl acetate; 3-methyl-2-butenyl acetate; (E)-2-hexenyl acetate; (E)- and (Z)-3-hexenyl acetate; octyl acetate; 3-octyl acetate; 1-octen-3-yl acetate; ethyl butyrate; butyl butyrate, isoamyl butyrate; hexyl butyrate; (E)- and (Z)-3-hexenyl isobutyrate; hexyl crotonate; ethyl isovalerate; ethyl 2-methylpentanoate; ethyl hexanoate; allyl hexanoate; ethyl heptanoate; allyl heptanoate; ethyl octanoate; ethyl (E,Z)-2,4-decadienoate; methyl 2-octynoate; methyl 2-nonynoate; allyl 2-isoamyloxyacetate; methyl 3,7-dimethyl-2,6-octadienoate;
of acyclic terpene alcohols, such as, for example, citronellol; geraniol; nerol; linalool; lavandulol; nerolidol; farnesol; tetrahydrolinalool; tetrahydrogeraniol; 2,6-dimethyl-7-octen-2-ol; 2,6-dimethyloctan-2-ol; 2-methyl-6-methylene-7-octen-2-ol; 2,6-dimethyl-5,7-octadien-2-ol; 2,6-dimethyl-3,5-octatrien-2-ol; 3,7-dimethyl-4,6-octadien-3-ol; 3,7-dimethyl-1,5,7-octatrien-3-ol; 2,6-dimethyl-2,5,7-octatrien-1-ol; and formates, acetates, propionates, isobutyrates, butyrates, isovalerates, pentanoates, hexanoates, crotonates, tiglinates, 3-methyl-2-butenoates thereof;
of acyclic terpene aldehydes and ketones, such as, for example, geranial; neral; citronellal; 7-hydroxy-3,7-dimethyloctanal; 7-methoxy-3,7-dimethyloctanal; 2,6,10-trimethyl-9-undecenal; geranylacetone; and the dimethyl and diethyl acetals of geranial, neral, 7-hydroxy-3,7-dimethyloctanal;
of cyclic terpene alcohols, such as, for example, menthol; isopulegol; alpha-terpineol; terpineol-4; menthan-8-ol; menthan-1-ol; menthan-7-ol; borneol; isoborneol; linalool oxide; nopol; cedrol; ambrinol; vetiverol; guaiol; and formates, acetates, propionates, isobutyrates, butyrates, isovalerates, pentanoates, hexanoates, crotonates, tiglinates, 3-methyl-2-butenoates thereof;
of cyclic terpene aldehydes and ketones, such as, for example, menthone; isomenthone; 8-mercaptomenthan-3-one; carvone; camphor; fenchone; alpha-ionone; beta-ionone; alpha-n-methylionone; beta-n-methylionone; alpha-isomethylionone; beta-isomethylionone; alpha-irone; alpha-damascone; beta-damascone; beta-damascenone; delta-damascone; gamma-damascone; 1-(2,4,4-trimethyl-2-cyclohexen-1-yl)-2-buten-1-one; 1,3,4,6,7,8a-hexahydro-1,1,5,5-tetramethyl-2H-2,4a-methanonaphthalen-8(5H)-one; nootkatone; dihydronootkatone; alpha-sinensal; beta-sinensal; acetylated cedarwood oil (methyl cedryl ketone);
of cyclic alcohols, such as, for example, 4-tert-butylcyclohexanol; 3,3,5-trimethylcyclohexanol; 3-isocamphylcyclohexanol; 2,6,9-trimethyl-Z2,Z5,E9-cyclododecatrien-1-ol; 2-isobutyl-4-methyltetrahydro-2H-pyran-4-ol;
of cycloaliphatic alcohols, such as, for example, alpha-3,3-trimethyl-cyclohexylmethanol; 2-methyl-4-(2,2,3-trimethyl-3-cyclopent-1-yl)butanol; 2-methyl-4-(2,2,3-trimethyl-3-cyclopent-1-yl)-2-buten-1-ol; 2-ethyl-4-(2,2,3-trimethyl-3-cyclopent-1-yl)-2-buten-1-ol; 3-methyl-5-(2,2,3-trimethyl-3-cyclopent-1-yl)-pentan-2-ol; 3-methyl-5-(2,2,3-trimethyl-3-cyclopent-1-yl)-4-penten-2-ol; 3,3-dimethyl-5-(2,2,3-trimethyl-3-cyclopent-1-yl)-4-penten-2-ol; 1-(2,2,6-trimethylcyclohexyl)pentan-3-ol; 1-(2,2,6-trimethylcyclohexyl)hexan-3-ol;
of cyclic and cycloaliphatic ethers, such as, for example, cineol; cedryl methyl ether; cyclododecyl methyl ether; (ethoxymethoxy)cyclododecane; alpha-cedrene epoxide; 3a,6,6,9a-tetramethyldodecahydronaphtho[2,1-b]furan; 3a-ethyl-6,6,9a-trimethyldodecahydronaphtho[2,1-b]furan; 1,5,9-trimethyl-13-oxabicyclo[10.1.0]trideca-4,8-diene; rose oxide; 2-(2,4-dimethyl-3-cyclohexen-1-yl)-5-methyl-5-(1-methylpropyl)-1,3-dioxane;
of cyclic ketones, such as, for example, 4-tert-butylcyclohexanone; 2,2,5-trimethyl-5-pentylcyclopentanone; 2-heptylcyclopentanone; 2-pentylcyclopentanone; 2-hydroxy-3-methyl-2-cyclopenten-1-one; 3-methyl-cis-2-penten-1-yl-2-cyclopenten-1-one; 3-methyl-2-pentyl-2-cyclopenten-1-one; 3-methyl-4-cyclopentadecenone; 3-methyl-5-cyclopentadecenone; 3-methylcyclopentadecanone; 4-(1-ethoxyvinyl)-3,3,5,5-tetramethylcyclohexanone; 4-tert-pentylcyclohexanone; 5-cyclohexadecen-1-one; 6,7-dihydro-1,1,2,3,3-pentamethyl-4(5H)-indanone; 5-cyclohexadecen-1-one; 8-cyclohexadecen-1-one; 9-cycloheptadecen-1-one; cyclopentadecanone;
of cycloaliphatic aldehydes, such as, for example, 2,4-dimethyl-3-cyclo-hexenecarbaldehyde; 2-methyl-4-(2,2,6-trimethylcyclohexen-1-yl)-2-butenal; 4-(4-hydroxy-4-methylpentyl)-3-cyclohexenecarbaldehyde; 4-(4-methyl-3-penten-1-yl)-3-cyclohexenecarbaldehyde;
of cycloaliphatic ketones, such as, for example, 1-(3,3-dimethylcyclohexyl)-4-penten-1-one; 1-(5,5-d imethyl-1-cyclohexen-1-yl)-4-penten-1-one; 2,3,8,8-tetramethyl-1,2,3,4,5,6,7,8-octahydro-2-naphthalenyl methyl ketone; methyl-2,6,10-trimethyl-2,5,9-cyclododecatrienyl ketone; tert-butyl 2,4-dimethyl-3-cyclohexen-1-yl ketone;
of esters of cyclic alcohols such as, for example, 2-tert-butylcyclohexyl acetate; 4-tert-butylcyclohexyl acetate; 2-tert-pentylcyclohexyl acetate; 4-tert-pentylcyclohexyl acetate; decahydro-2-naphthyl acetate; 3-pentyltetrahydro-2H-pyran-4-yl acetate; decahydro-2,5,5,8a-tetramethyl-2-naphthyl acetate; 4,7-methano-3a,4,5,6,7,7a-hexahydro-5 or 6-indenyl acetate; 4,7-methano-3a,4,5,6,7,7a-hexahydro-5 or 6- indenyl propionate; 4,7-methano-3a,4,5,6,7,7a-hexahydro-5 or 6-indenyl isobutyrate; 4,7-methanooctahydro-5 or 6-indenyl acetate;
of esters of cycloaliphatic carboxylic acids, such as, for example, allyl 3-cyclohexyl-propionate; allyl cyclohexyloxyacetate; methyl dihydrojasmonate; methyl jasmonate; methyl 2-hexyl-3-oxocyclopentanecarboxylate; ethyl 2-ethyl-6,6-dimethyl-2-cyclohexenecarboxylate; ethyl 2,3,6,6-tetramethyl-2-cyclohexenecarboxylate; ethyl 2-methyl-1,3-dioxolan-2-acetate;
of aromatic hydrocarbons, such as, for example, styrene and diphenylmethane;
of araliphatic alcohols, such as, for example, benzyl alcohol; 1-phenylethyl alcohol; 2-phenylethyl alcohol; 3-phenylpropanol; 2-phenylpropanol; 2-phenoxyethanol; 2,2-dimethyl-3-phenylpropanol; 2,2-dimethyl-3-(3-methylphenyl)propanol; 1,1-dimethyl-2-phenylethyl alcohol; 1,1-dimethyl-3-phenylpropanol; 1-ethyl-1-methyl-3-phenylpropanol; 2-methyl-5-phenylpentanol; 3-methyl-5-phenylpentanol; 3-phenyl-2-propen-1-ol; 4-methoxybenzyl alcohol; 1-(4-isopropylphenyl)ethanol;
of esters of araliphatic alcohols and aliphatic carboxylic acids, such as, for example, benzyl acetate; benzyl propionate; benzyl isobutyrate; benzyl isovalerate; 2-phenylethyl acetate; 2-phenylethyl propionate; 2-phenylethyl isobutyrate; 2-phenylethyl isovalerate; 1-phenylethyl acetate; alpha-trichloromethylbenzyl acetate; alpha, alpha-dimethylphenylethyl acetate; alpha,alpha-dimethylphenylethyl butyrate; cinnamyl acetate; 2-phenoxyethyl isobutyrate; 4-methoxybenzyl acetate; of araliphatic ethers, such as, for example, 2-phenylethyl methyl ether; 2-phenylethyl isoamyl ether; 2-phenylethyl 1-ethoxyethyl ether; phenylacetaldehyde dimethyl acetal; phenylacetaldehyde diethyl acetal; hydratropaldehyde dimethyl acetal; phenylacetaldehyde glycerol acetal; 2,4,6-trimethyl-4-phenyl-1,3-dioxane; 4,4a,5,9b-tetrahydroindeno[1,2-d]-m-dioxin; 4,4a,5,9b-tetrahydro-2,4-dimethylindeno[1,2-d]-m-dioxin;
of aromatic and araliphatic aldehydes, such as, for example, benzaldehyde; phenylacetaldehyde; 3-phenylpropanal; hydratropaldehyde; 4-methylbenzaldehyde; 4-methylphenylacetaldehyde; 3-(4-ethylphenyl)-2,2-dimethylpropanal; 2-methyl-3-(4-isopropyl phenyl)propanal; 2-methyl-3-(4-tert-butylphenyl)propanal; 3-(4-tert-butylphenyl)propanal; cinnamaldehyde; alpha-butylcinnamaldehyde; alpha-amylcinnamaldehyde; alpha-hexylcinnamaldehyde; 3-methyl-5-phenylpentanal; 4-methoxybenzaldehyde; 4-hydroxy-3-methoxybenzaldehyde; 4-hydroxy-3-ethoxybenzaldehyde; 3,4-methylenedioxybenzaldehyde; 3,4-dimethoxy-benzaldehyde; 2-methyl-3-(4-methoxyphenyl)propanal; 2-methyl-3-(4-methylenedioxyphenyl)-propanal;
of aromatic and araliphatic ketones, such as, for example, acetophenone; 4-methylacetophenone; 4-methoxyacetophenone; 4-tert-butyl-2,6-dimethylacetophenone; 4-phenyl-2-butanone; 4-(4-hydroxyphenyl)-2-butanone; 1-(2-naphthalenyl)ethanone; benzophenone; 1,1,2,3,3,6-hexamethyl-5-indanyl methyl ketone; 6-tert-butyl-1,1-dimethyl-4-indanyl methyl ketone; 1-[2,3-dihydro-1,1,2,6-tetramethyl-3-(1-methylethyl)-1H-5-indenyl]ethanone; 5xe2x80x2,6xe2x80x2,7xe2x80x2,8xe2x80x2-tetrahydro-3xe2x80x2,5xe2x80x2,5xe2x80x2,6xe2x80x2,8xe2x80x2,8xe2x80x2-hexamethyl-2-acetonaphthone;
of aromatic and araliphatic carboxylic acids and esters thereof, such as, for example, benzoic acid; phenylacetic acid; methyl benzoate; ethyl benzoate; hexyl benzoate; benzyl benzoate; methyl phenylacetate; ethyl phenylacetate; geranyl phenylacetate; phenylethyl phenylacetate; methyl cinnamate; ethyl cinnamate; benzyl cinnamate; phenylethyl cinnamate; cinnamyl cinnamate; allyl phenoxyacetate; methyl salicylate; isoamyl salicylate; hexyl salicylate; cyclohexyl salicylate; cis-3-hexenyl salicylate; benzyl salicylate; phenylethyl salicylate; methyl 2,4-dihydroxy-3,6-dimethylbenzoate; ethyl 3-phenylglycidate; ethyl 3-methyl-3-phenylglycidate;
of nitrogen-containing aromatic compounds, such as, for example, 2,4,6-trinitro-1,3-dimethyl-5-tert-butylbenzene; 3,5-dinitro-2,6-dimethyl-4-tert-butylacetophenone; cinnamonitrile; 5-phenyl-3-methyl-2-pentenenitrile; 5-phenyl-3-methylpentanenitrile; methyl anthranilate; methyl N-methylanthranilate; Schiff bases of methyl anthranilate with 7-hydroxy-3,7-dimethyloctanal; 2-methyl-3-(4-tert-butylphenyl)propanal or 2,4-dimethyl-3-cyclohexenecarbaldehyde; 6-isopropylquinoline; 6-isobutylquinoline; 6-sec-butylquinoline; indole; skatole; 2-methoxy-3-isopropylpyrazine; 2-isobutyl-3-methoxypyrazine;
of phenols, phenyl ethers and phenyl esters, such as, for example, estragole; anethole; eugenole; eugenyl methyl ether; isoeugenole; isoeugenyl methyl ether; thymole; carvacrol; diphenyl ether; beta-naphthyl methyl ether; beta-naphthyl ethyl ether; beta-naphthyl isobutyl ether; 1,4-dimethoxybenzene; eugenyl acetate; 2-methoxy-4-methylphenol; 2-ethoxy-5-(1-propenyl)phenol; p-cresyl phenylacetate;
of heterocyclic compounds, such as, for example, 2,5-dimethyl-4-hydroxy-2H-furan-3-one; 2-ethyl-4-hydroxy-5-methyl-2H-furan-3-one; 3-hydroxy-2-methyl-4H-pyran-4-one; 2-ethyl-3-hydroxy-4H-pyran-4-one;
of lactones, such as, for example, 1,4-octanolide; 3-methyl-1,4-octanolide; 1,4-nonanolide; 1,4-decanolide; 8-decen-1,4-olide; 1,4-undecanolide; 1,4-dodecanolide; 1,5-decanolide; 1,5-dodecanolide; 1,15-pentadecanolide; cis- and trans-11-pentadecen-1,15-olide; cis- and trans-12-pentadecen-1,15-olide; 1,16-hexadecanolide; 9-hexadecen-1,16-olide; 10-oxa-1,16-hexadecanolide; 11-oxa-1,16-hexadecanolide; 12-oxa-1,16-hexadecanolide; ethylene 1,12-dodecanedioate; ethylene 1,13-tridecanedioate; coumarin; 2,3-dihydrocoumarin; octahydrocoumarin.
The care agents according to the present invention generally comprise 0.1 to 20 parts by weight, preferably 0.5 to 2 parts by weight, more preferably 0.8 to 1.2 parts by weight, of the perfume component.
The care agents according to the present invention further comprise a washing-active surfactant component. The washing-active surfactant component can consist of one or more, preferably 3 to 5, surfactants.
Washing-active surfactants for the care agents according to the present invention are known per se (Andreas Domsch, Die kosmetischen Prxc3xa4parate, Band II [Cosmetic Preparations, Volume II]).
By way of example, mention may be made of the following washing-active surfactants.
This class of substance includes secondary alkanesulfonates, e.g. Sodium C14-17 alkyl Sec Sulfonate, alkyl sulfates e.g. Sodium Lauryl Sulfate, alkyl ether sulfates, lauryl ether sulfates e.g. Sodium Laureth Sulfate, olefinsulfonates, e.g. Sodium Olefin C14-16 Sulfonate, alkylamide ether sulfates, e.g. TEA-PEG-3 Cocamide Sulfate, acyl isethionates e.g. Sodium Cocoyl Isethionate, acyl glutamates e.g. Lauroyl Glutamate, alkyl ether carboxylates e.g. Trideceth-7 Carboxylic Acid, methyl taurides e.g. Sodium Lauroyl Taurate, sarcosides, e.g. Ammonium Cocoyl Sarcosinate, sulfosuccinates, e.g. Disodium PEG-4 Cocamido MIPA-Sulfosuccinate, protein fatty acid condensates e.g. Potassium Cocoyl Hydrolyzed Collagen.
This class of substance includes alkylbetaines, e.g. Lauryl Betaine, alkylamidobetaines e.g. Cocamidopropyl Betaine, sulfobetaines e.g. Cocamidopropyl Hydroxysultaine, acetates and diacetates, e.g. Disodium Cocoamphodiacetate, imidazolines, e.g. Amphonyl, propionates. e.g. Sodium Cocoamphopropionate.
This class of substance includes ethoxylated castor oil derivatives, e.g. PEG-20 Hydrogenated Castor Oil, polysorbates, e.g. Polysorbate 80, glycerol fatty acid ester ethoxylates, e.g. PEG 20 Glyceryl Isostearate, sugar surfactants, e.g. capryl/caprylyl glucosides, amine oxides, e.g. Cocamine Oxide.
The care agents according to the present invention generally contains 0.1 to 50 parts by weight, preferably 1 to 20 parts by weight, more preferably 5 to 10 parts by weight, of the washing-active surfactant component.
The care agents according to the present invention comprise water as a further component.
Furthermore, the care agents according to the present invention can also contain electrolytes.
Examples of electrolytes for the care agents according to the present invention which may be mentioned are: sodium chloride, ammonium chloride and magnesium chloride.
The care agents according to the present invention can generally comprise 0.1 to 5 parts by weight, preferably 0.3 to 3 parts by weight, particularly preferably 0.5 to 2 parts by weight, of electrolytes.
The care agents according to the present invention can be prepared as follows:
The gellan gum is dissolved in water at about 70xc2x0 C., then cooled to about 55xc2x0 C., and the perfume component is added. The solution is homogenized using a high-speed stirrer and cooled to about 45xc2x0 C. with stirring. The solution is stirred throughout the entire preparation. A dosing pump is used to pump the solution into an ice-cooled aqueous calcium bath with a calcium ion concentration of about 6 mmol.
The washed matrix particles are added to the surfactant formulation.
The care agents prepared by this process contain matrix particles with a spherical diameter of from 3 to 5 mm. The shape of the particles is uniformly round. The particles have an average degree of hardness and can be destroyed without targeted mechanical action, as arises, for example, during the showering or washing operation.
The care agents according to the present invention can be used in the field of body-cleansing and in technical cleaners, e.g. household cleaners.
Surprisingly, the care agents according to the present invention based on matrix particles have a high retention behavior for (surfactant components), perfume oils and other active ingredients.
It is readily possible to also add solid components, for example, color pigments, to the matrix particles.
One advantage of the care agents according to the present invention based on matrix particles is their simple formulation make-up and an uncomplicated preparation process. The uncomplicated arrangement of apparatus can, therefore, be transferred easily to production scale.
The present invention displays the following further unexpected advantages:
Care agents with the matrix particles according to the present invention do not tend toward xe2x80x9cbleedingxe2x80x9d of the filling material
Care agents based on the matrix particles have high odor stability in surfactant systems
Care agents based on the matrix particles have high chemical stability and adhesive effect
The care agents according to the present invention based on matrix particles permit a targeted use of the surfactant and perfume components and do not have a negative impact on the environment as a result of a high concentration.
The invention is further illustrated but is not intended to be limited by the following examples in which all parts and percentages are by weight unless otherwise specified.